硒蛋白SELENOI通过PI3K/AKT/mTOR信号通路促进胃癌增殖、迁移和侵袭OA
Selenoprotein SELENOI promotes the proliferation,migration and invasion of gastric cancer through the PI3K/AKT/mTOR signaling pathway
目的·探讨硒蛋白I(SELENOI,即乙醇胺磷酸转移酶1)对胃癌细胞增殖、迁移与侵袭的调控作用及其分子机制.方法·利用蛋白质印迹法(Western blotting)和实时荧光定量PCR(quantitative real-time PCR,qPCR)检测SELENOI在胃黏膜上皮细胞及胃癌细胞系中的表达水平;收集2010-2015年上海交通大学医学院附属仁济医院胃肠外科100例胃腺癌患者的临床病理资料,分析SELENOI表达水平与患者预后的关联;基于癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库分析SELENOI在胃腺癌肿瘤组织和正常组织中的表达.筛选SELENOI高、低表达组间的差异基因,并进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析,筛选SELENOI相关信号通路,同时通过 qPCR和蛋白质印迹法验证 SELENOI与该通路的调控关联;分别采用靶向 SELENOI的小干扰 RNA(small interfering RNA,siRNA)、SELENOI过表达质粒单独或联合哺乳动物雷帕霉素靶蛋白(mechanistic target of rapamycin,mTOR)抑制剂雷帕霉素处理HGC-27、NUGC-3胃癌细胞,通过细胞计数试剂盒-8(CCK-8)实验检测细胞增殖能力,Transwell实验检测细胞迁移、侵袭能力.结果·生物信息学分析发现SELENOI在胃癌组织中显著高表达(P<0.05);100例胃腺癌患者Kaplan-Meier生存分析显示,SELENOI高表达患者预后更差(P=0.022);qPCR和蛋白质印迹法结果证实,SELENOI在HGC-27、NUGC-3、AGS、MKN45胃癌细胞系中的表达水平显著高于胃黏膜上皮细胞GES-1(均P<0.05);KEGG通路富集分析以及后续分子验证实验证实,胃癌中SELENOI可促进磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶 B(protein kinase B,AKT)/mTOR 信号通路活化;CCK-8 与 Transwell 实验结果表明,SELENOI可通过激活PI3K/AKT/mTOR信号通路增强胃癌细胞的增殖、迁移和侵袭能力,而雷帕霉素可显著逆转该促癌效应.结论·SELENOI在胃癌组织及胃癌细胞中呈高表达状态;SELENOI可通过激活PI3K/AKT/mTOR信号通路促进胃癌细胞增殖、迁移及侵袭.
Objective·To investigate the regulatory role of the selenoprotein SELENOI(ethanolamine phosphotransferase 1,EPT1)in the proliferation,migration and invasion of gastric cancer cells,as well as its molecular mechanism.Methods·The expression levels of SELENOI in gastric mucosal epithelial cells and gastric cancer cell lines were detected using Western blotting and quantitative real-time PCR(qPCR).The clinical pathological data of 100 patients who underwent surgery at the Department of Gastrointestinal Surgery of Renji Hospital,Shanghai Jiao Tong University School of Medicine,from 2010 to 2015 were collected,and the association between SELENOI expression levels and patient prognosis was analyzed.The expression of SELENOI in gastric adenocarcinoma tumor tissues and normal tissues was analyzed based on The Cancer Genome Atlas(TCGA)database.The differentially expressed genes between the SELENOI high-and low-expression groups were screened,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was performed to select SELENOI-related signaling pathways.The regulatory association of SELENOI with this pathway was verified by qPCR and Western blotting.HGC-27 and NUGC-3 gastric cancer cells were treated with SELENOI-targeted small interfering RNA(siRNA),SELENOI overexpression plasmid alone,or in combination with the mechanistic target of rapamycin(mTOR)inhibitor rapamycin.The cell proliferation ability was detected by CCK-8 assay,and cell migration and invasion abilities were detected by Transwell assay.Results·Bioinformatics analysis revealed that SELENOI was significantly highly expressed in gastric cancer tissues(P<0.05).Kaplan-Meier survival analysis of 100 patients with gastric adenocarcinoma showed that patients with high SELENOI expression had worse prognosis(P=0.022).qPCR and Western blotting results confirmed that the expression levels of SELENOI in HGC-27,NUGC-3,AGS,and MKN45 gastric cancer cell lines were significantly higher than those in gastric mucosal epithelial cells GES-1(all P<0.05).KEGG pathway enrichment analysis and subsequent molecular validation experiments confirmed that SELENOI promoted the activation of the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mTOR signaling pathway in gastric cancer.CCK-8 and Transwell assay results showed that SELENOI enhanced the proliferation,migration and invasion abilities of gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway,and rapamycin significantly reversed this tumor-promoting effect.Conclusion·SELENOI is highly expressed in gastric cancer tissues and cells.SELENOI can promote the proliferation,migration and invasion of gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway.
王铭杰;倪博;夏翔;张子臻
上海交通大学医学院附属仁济医院胃肠外科,上海 200127上海交通大学医学院附属仁济医院胃肠外科,上海 200127上海交通大学医学院附属仁济医院胃肠外科,上海 200127上海交通大学医学院附属仁济医院胃肠外科,上海 200127
医药卫生
硒蛋白I磷脂酰肌醇3-激酶蛋白激酶B哺乳动物雷帕霉素靶蛋白胃癌
SELENOIphosphatidylinositol 3-kinase(PI3K)protein kinase B(AKT)mechanistic target of rapamycin(mTOR)gastric cancer
《上海交通大学学报（医学版）》 2026 (8)
1015-1025,11
国家自然科学基金(82103510,82303428,82473077,82473065)癌基因与相关基因国家重点实验室科研项目(课题)(KF2205-93)上海市卫生健康委员会医学新技术研究与转化种子计划项目(2024ZZ2020)上海市科学技术委员会三维电子腹腔内窥镜(微创外科领域)应用示范项目(25SF1905600)中央高校基本科研业务费专项资金(YG2026ZD15)仁济医院临床研究中心横向课题(IIT-2023-0001). National Natural Science Foundation of China(82103510,82303428,82473077,82473065)Research Project of the State Key Laboratory of Oncogenes and Related Genes(KF2205-93)Medical New Technology Research and Translation Seed Program of the Shanghai Municipal Health Commission(2024ZZ2020)Application Demonstration Project of Three-Dimensional Electronic Laparoscope in Minimally Invasive Surgery of the Science and Technology Commission of Shanghai Municipality(25SF1905600)Fundamental Research Funds for the Central Universities(YG2026ZD15)Horizontal Research Project of the Clinical Research Center,Renji Hospital(IIT-2023-0001).
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